Journal of Bioanalysis & Biomedicine - Open Access

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Research Article
OPEN ACCESS Freely available online
doi:10.4172/1948-593X.1000024

JBABM/Vol.2 Issue 3

Pharmacokinetics of Gentamicin and its Interaction with

Paracetamol after i.v. Administration in Buffalo Calves
(Bubalus bubalis)
Baxla S.L1, Kumar M2, Jayachandran C2, Roy B.K1*and Kumari A2
1
2

Department of Pharmacology &Toxicology, College of Veterinary Science & A.H, B.A.U., Ranchi-834006, India
Department of Pharmacology & Toxicology, Bihar Veterinary College, R.A.U., Patna-800014, India

Abstract
The disposition kinetics was conducted in five healthy female buffalo calves following single i.v dose (5mg/kg) of
gentamicin alone and with paracetamol (40mg/kg,i.v.). The study revealed that the plasma concentrations of gentamicin
were significantly higher when it was given with paracetamol as compared to alone between 0.042 to 0.333 hrs and at 24
hrs. Serum concentrations of gentamicin was detected for longer period (48 hrs) in urine in both groups of experimental
animals. In case of urine drug attained its peak level at the same time interval (1.5 hrs) in both groups with the
concentration of 83.423.14 g/ml after alone and 545.125.85 g/ml with paracetamol administration. The extrapolated
zero time plasma concentrations during distribution phase (A) and theoretical zero time plasma concentrations (Cp)
were significantly (p< 0.01) higher 34.482.35 and 39.032.40 g/ml respectively. Also significantly higher distribution
rate constant () of 1.9350.1119 h-1 and lower distribution half life (t1/2 ) of 0.360.02 hrs were observed, when
gentamicin was given with paracetamol. Elimination half life (t1/2 ) of 6.670.11 hrs was not significantly higher when
gentamicin was given with paracetamol. AUC (62.162.82 mg/L.hrs) was significantly (p< 0.05) higher while MRT
(6.930.36hrs) was not significantly higher when gentamicin was given with paracetamol. The values of K12, K21 and
Kel were calculated to be 1.0880.111 h-1, 0.3230.028 h-1and 0.6280.024 h-1 respectively when gentamicin was given
concurrently with paracetamol. T~P (5.040.16) was significantly (p< 0.01) higher, while Vdarea (0.780.03L/Kg) and
ClB (1.350.06 ml/Kg/min) were not significantly higher when gentamicin was given with paracetamol. The present
investigation established that both gentamicin and paracetamol interacted with altered their kinetic behaviour. The
combination with paracetamol may be benificial because paracetamol reduced the maintenance doses of gentamicin
which may be much advantageous in the field of veterinary practice in the dose of 5 mg/kg daily by systemic route and
36 hourly when given with paracetamol in urinary tract infection.

Keywords: Pharmacokinetics; Gentamicin; Paracetamol; Interaction;

Intravenous; Buffalo calves
Abbreviations: A: Zero time plasma drug concentration during

distribution phase; B: Zero time plasma drug concentration

during elimination phase; Cp: theoretical zero time plasma drug
concentration; : distribution rate constant; t: distribution half
life; : Elimination rate constant; t: elimination half life; AUC: Area
under curve; AUMC: Area under first moment curve; MRT: Mean
residence time; K12: Rate constants for drug transfer from central to
peripheral compartment; K21: Rate constants for drug transfer from
peripheral to central compartment; Kel: Rate constants for elimination
of drug from central compartment; Fc: Fraction of drug in central
compartment; T~P: Tissue to plasma concentration; VdC: Volume of
the central compartment; VdSS: Apparent volume of distribution at
steady state; VdB: Extrapolated volume of distribution; Vdarea: Volume
of distribution by area; ClB: Total body clearance

for treating febrile conditions. Paracetamol indirectly blocks Coxenzyme and that this blockade is ineffective in the presence of
peroxides. Cox also produces thromboxanes, which aid in the blood
clotting, aspirin reduces blood clotting, but paracetamol does not.
Thus paracetamol is effective in the central nervous system and in
endothelial cells. It does not affect function of platelets and clotting
factors and is less gastrointestinal irritant. It is a suitable substitute
for aspirin for its antipyretic or analgesic actions.
Although pharmacokinetic study of gentamicin have been
conducted in many species of animals,it seems little work has been
done on kinetic interaction of gentamicin with NSAIDs in buffalo
calves, particularly on the interaction of gentamicin with paracetamol.
Therefore, the present experiment aims on the pharmacokinetic
studies of gentamicin and its interaction with paracetamol after i.v.
administration in buffalo calves.

Introduction

Paracetamol, a potent antipyretic agent, having analgesic and

anti-inflammatory properties and is widely used in man and animals

Received May 20, 2010; Accepted June 28, 2010; Published June 28, 2010
Citation: Baxla SL, Kumar M, Jayachandran C, Roy BK, Kumari A (2010)
Pharmacokinetics of Gentamicin and its Interaction with Paracetamol after i.v.
Administration in Buffalo Calves (Bubalus bubalis). J Bioanal Biomed 2: 065-068.
doi:10.4172/1948-593X.1000024
Copyright: 2010 Baxla SL, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.

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*Corresponding authors: Dr. B.K.Roy, Uni. Prof. & Chairman, Department

of Pharmacology &Toxicology, College of Veterinary Science & A.H, B.A.U.,
Ranchi-834006, India, Tel: +91-651-2450759(o); Fax: +91-651-2450759(o);
E-mail: roybk2001@yahoo.co.in

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Gentamicin, a broad spectrum aminoglycoside is widely used in

infectious diseases of animals. It is used to treat various infections
caused by aerobic Gram negative bacteria such as E.coli, Salmonella,
Klebsiella, Proteus, Haemophilus, Pasteurella, Compylobactor and
Pseudomonas. It binds to 30s ribosomal subunit; however, it also
appear to bind to several sites on the 50s ribosomal subunit. It is
therapeutically used in cases of urinary tract infections, bacteremia,
infected burns, osteomyelitis, pneumonia, peritonitis and otitis.

Citation: Baxla SL, Kumar M, Jayachandran C, Roy BK, Kumari A (2010) Pharmacokinetics of Gentamicin and its Interaction with Paracetamol after
i.v. Administration in Buffalo Calves (Bubalus bubalis). J Bioanal Biomed 2: 065-068. doi:10.4172/1948-593X.1000024

Animals
Five clinically healthy female buffalo calves of non descript breed,
of 12 to 18 months age, weighing between 102 to180 kg were used
in this experiment. The calves were procured by cattle breeding farm
of Bihar Veterinary College Patna. They were maintained exclusively
on grazing and also supplementary concentrate feeding was made
for them with the provision of night shelter. Water was given ad
libitum throughout the period of investigation. The protocol of the
experiment was approved by Institutional Animal Ethics Committee.

Drugs used
Progenta, an injectable commercial preparation containing
gentamicin in concentration of 40 mg/ml marketed by Vetsfarma
(@5mg/kg b. wt) and Paracetol-vet-an injectable commercial
preparation containing paracetamol in concentration of 150 mg/ml
marketed by Sarabhai Zydus (@40mg/kg b.wt) were administered
separately in each of five buffalo calves by i.v.route.An interval of 15
days was allowed to elapse before administration of next dose of the
drug. After conducting kinetic study of gentamicin and paracetamol
by i.v. route separately, both the drugs were administered together at
above dose rate in each animal by i.v. route to identify the interaction
if any.

Collection of samples
The samples of various biological fluids (plasma & urine) were
collected at 0.042, 0.083, 0.167, 0.25, 0.333, 0.50, 0.75 min and 1,
1.5, 2, 3, 4, 5, 6, 8, 10, 12 and 24 hrs. The samples of urine were
collected further upto 48 hrs (30, 36 and 48hrs).

Blood
Blood samples were collected in sterilized centrifuge tubes
containing appropriate amount of sodium oxalate by vene-puncture
with disposable 18G needle at above noted time intervals after drug
administration. The blood samples were centrifuged at 3000 rpm
for 10 min for the separation of plasma and kept under refrigeration
until assay was carried out.

Urine
Urine samples were collected in a sterile test tubes for analysis by
introducing a sterile Foley`s balloon catheter (No. 12) lubricated with
glycerine through urethra into urinary bladder of the experimental
buffalo calves with the aid of a flexible metal probe. The balloon of
the catheter was inflated by injecting 25 -30 ml of water through a
syringe to keep the catheter in position. The opening of the catheter
was blocked with a pressure clip to check dripping of urine.

Analysis of gentamicin
Estimation of gentamicin was carried out by microbiological
assay technique (cylinder plate diffusion method) using Bacillus
subtilis (ATCC 6633) as the test organism (Grove and Randell, 1955;
Orsini et al., 1985). The culture of Bacillus subtilis was obtained from
National Collection of Industrial Microorganism (NCIM), Division of
Biochemical Sciences, National Chemical Laboratory, Pune- 8. The
organism was grown on the slant of culture tube containing nutrient
agar slants at 37C for overnight. The organism was transferred
weekly to fresh media to maintain its normal activity. Gentamicin was
diluted in sterile glass distilled water to have different strengths viz.,

Analysis of paracetamol
Paracetamol was estimated by spectrophotometric method
(Archer and Richardson, 1980; Omer and Mohammod, 1984).
Paracetamol was diluted in glass distilled water to different strength
viz. 1000, 500, 250, 100, 50 and 20 l/ml. From each standard
solution, 0.1ml was added to a sterile vials containing 0.9ml of plasma
or urine collected prior to drug administration. These standards were
simultaneously used along with test samples for the determination of
drug concentrations in biological fluids.

Pharmacokinetic analysis
The pharmacokinetic parameters of gentamicin and paracetamol
were calculated after its single i.v. administration from semi log plot
of plasma drug concentration versus time curve. The experimental
data was analyzed by using two compartment open model
(Gibaldi and Perrier, 1975; Baggot, 1974; Notari, 1987). For a two
compartment model, the concentration of the drug in plasma at any
time is obtained from the formula.
Cp =Ac-t + Be-t
Where, Cp is the drug concentration in plasma at timet. A, the
zero time concentration of the drug in plasma and , the regression
coefficient (distribution rate constant) for distribution phase
were calculated by the method of residual yield. B, the zero time
concentration of the drug in plasma and , the regression coefficient
(elimination rate constant) for elimination phase were calculated by
the method of least squares.

Calculation of dosage regimen

Dosage regimen was calculated for antimicrobial agent to
maintain minimum inhibitory concentration (MIC) in plasma at
desired dosage intervals. Leroy et al. (1978) reported the therapeutic
plasma levels (MICs) of gentamicin to be 1-4 g/ml. Hence, in the
present study, dosage regimen of gentamicin were calculated at 1,2
and 4 g/ml levels for the dosage intervals () 8 and 12 hrs using the
formulas (Saini and Shrivastva, (1997).
D* = Cp (min). Vdarea (e)
Do = Cp (min). Vdarea (e-1)
Where D* = Loading or priming dose (mg/kg), Do= Maintenance
dose (mg/kg), Cp (min) = Desired minimum plasma concentration
(g/ml), = Dosage interval (hrs), and Vdarea were obtained from
kinetic study.

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80, 40, 20, 10, 5, 2, 1 and 0.5 g/ml. From each of these solutions,
0.1ml was taken with the aid of micropipette and added to sterile
vials containing 0.9ml of plasma or urine collected prior to drug
administration. This yielded drug standards of 8, 4, 2, 1, 0.5, 0.2,
0.1 and 0.05 g/ml in the respective biological fluids. These standard
samples were stored in refrigerator and used simultaneously with test
samples in assay plates for obtaining standard curve. 50 microlitres of
each standard solution of various strength as well as test samples of
the drug were poured in separate porcelin cylinder in the assay plate.
The mean diameters of the bacterial zones of inhibition produced by
the standard as well as test samples of the drug were measured. The
standard curve was plotted from the measure of zone of inhibition
against each concentration of the drug on a semi log scale. With
the help of this standard curve and measured zone of inhibition f
different test sample, concentrations of drug in test samples were
estimated.

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Materials and Methods

Journal of Bioanalysis & Biomedicine - Open Access

Research Article
OPEN ACCESS Freely available online
doi:10.4172/1948-593X.1000024

www.omicsonline.org

JBABM/Vol.2 Issue 3

Results

Comparative Semilogerithmic plasma concentrations

of Gentamicin alone and with paracetamol

Concentration (g/ml)

100
Gentamicin
alone
Gentamicin+
Paracetamol

10

0.

Time (h)

Figure 1:

24

12

10

Concentration (g/ml)

100
10
1
0.1

0.01

Time (h)

Figure 2:
Kinetic parameters (unit)
Gentamicin alone
Gentamicin + paracetamol
A (g/ml)
11.08 0.86
34.48 2.35 **
NS
B (g/ml)
4.81 0.31
4.41 0.21
Cp (g/ml)
15.89 0.85
39.03 2.40 **
(h-1)
1.058 0.10
1.935 0.119 *
t1/2 (hrs)
0.69 0.08
0.36 0.02 *
-1
NS
(h )
0.139 0.008
0.104 0.01
t1/2 (hrs)
5.05 0.30
6.67 0.11 NS
AUC (mg/L.hrs)
45.90 3.10
62.16 2.82 *
AUMC (mg/L.h2)
268.2 27.86
433.1 28.63*
NS
MRT (hrs)
5.79 0.34
6.97 0.36
-1
K12 (h )
0.425 0.054
1.088 0.111 *
-1
NS
K21 (h )
0.421 0.42
0.323 0.028
-1
Kel (h )
0.351 0.023
0.628 0.024 **
Fc
0.40 0.02
0.17 0.01 **
T~P
1.54 0.16
5.04 0.16 **
VdC (L/kg)
0.32 0.02
0.13 0.01 **
VdB (L/kg)
1.05 0.06
1.11 0.06 *
NS
Vdarea (L/kg)
0.80 0.04
0.78 0.03
NS
VdSS (L/kg)
0.64 0.04
0.56 0.04
NS
ClB (ml/kg/min)
1.86 0.16
1.35 0.06
NS
Non significant
* p < 0.05
** p < 0.01

Table 1: Mean kinetic parameters of gentamicin alone (5mg/kg) and with

paracetamol (40mg/kg) after i.v.administration in buffalo calves.

was given alone while lower D*s of 3.6 & 7.1 and DOs of 2.0 and 4.0
mg/kg is required when gentamicin is given with paracetamol. Hence,
shorter of 8hrs can be recommended.

Discussion
Distribution in plasma and urinary excretion as well as
different kinetic parameters of gentamicin when given alone and
in combination with paracetamol following i.v.administration
interacted with one another. Paracetamol influenced the kinetics
of gentamicin.Serum concentrations of gentamicin were detected
upto 24hrs when gentamicin was given alone and when given
with paracetamol whereas gentamicin was detectable only upto
6hrs in febrile and afebrile conditions of goat @5mg\kg after i.v.
administration (Ahmad et al., 1994).The plasma concentrations of
gentamicin were found to be significantly higher initially (0.042 to
0.333 hrs) when it was given in combination with paracetamol as
compared to its alone administration. In case of urine, concentration
of gentamicin persisted for a longer period (36hrs) when it was given
with paracetamol. The t1\2 was significantly lower (0.360.02
hrs) when gentamicin was given with paracetamol as compared to
its alone administration (0.690.08). The above findings of this
experiment clearly indicated that paracetamol influenced the rate of
distribution of gentamicin and gentamicin was distributed at a faster

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1.

04
0. 2
08
0. 3
16
7
0.
2
0. 5
33
3
0.
5
0.
75

0.1

Gentamicin
Alone
Gentamicin+
Paracetamol

1000

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The calculated loading doses (D*s) for treating mild, moderate and
severe systemic infections (Cpmin =1, 2 and 4 g/ml respectively)
when gentamicin was given alone and with paracetamol, did not
differ significantly while maintenance doses (DOs) were noted to be
significantly lower (p< 0.05) for the dosage interval () of both 8
and 12 hrs when gentamicin was given along with paracetamol as
compared to its alone administration. For treating moderate (Cpmin
= 2g/ml) and severe (Cpmin = 4g/ml) infections at of 8hrs D*s of
4.9 and 9.9, DOs of 3.3 and 6.7 mg/kg were needed when gentamicin

Comparative semilogerithmic urine concentration

of gentamicin alone and with paracetamol

0.
0
0. 42
0
0. 83
16
0 7
0. .25
33
3
0
0. .5
75
1
1.
5
2
3
4
5
6
8
10
12
24
30
36
48

Plasma concentrations of gentamicin were found to be significantly

higher when it was given with paracetamol as compared to its alone
administration from 0.042 to 0.333 hrs and 24 hrs. (Figure 1). The
therapeutic concentration ( 2 g/ml) was maintained upto 6 hrs in
both the groups. Serum concentrations of gentamicin were detected
upto 24 hrs in both the groups of buffalo calves. Concentrations of
the gentamicin in urine were significantly higher from 0.083 to 24
hrs when gentamicin was given with paracetamol (Figure 2). The drug
attained its peak level in urine at the same time interval of 1.5 hrs
in both the groups with a concentration of 83.423.17 g/ml when
gentamicin was given alone and 545.125.85 g/ml with paracetamol.
The mean therapeutic concentration in urine (2 g/ml) was
maintained up to 24hrs when gentamicin was given alone and 36hrs
when given with paracetamol. The values of A, and Cp were noted to
be significantly higher (p < 0.01) in case of combined administration
as compared to alone administration of gentamicin. Area under curve
(AUC), area under first moment curve (AUMC) and micro rate constant
from central to peripheral compartment (K12) were significantly
higher (p < 0.01) when gentamicin was given with paracetamol
as compared to its alone administration. Rate constant of drug
elimination from central compartment (Kel), and approximate tissue
to plasma concentration ratio (T~P) were noted to be 0.6280.024
and 5.040.16 for gentamicin when it was given with paracetamol.
These values were significantly higher (p < 0.01) as compared to
alone administration of gentamicin. The value of distribution half-life
(t1/2) was significantly (p<0.05) lower (0.360.02hrs) in combined
administration of gentamicin as compared to its alonee administration
(0.690.08hrs). The extrapolated zero time concentration during
elimination phase (B),elimination rate constant (), elimination half
life (t1\2), mean residence time (MRT), rate constant of transfer of
drug from peripheral to central compartment (K21) and Vdarea did
not differ significantly between both the groups (Table 1).

Citation: Baxla SL, Kumar M, Jayachandran C, Roy BK, Kumari A (2010) Pharmacokinetics of Gentamicin and its Interaction with Paracetamol after
i.v. Administration in Buffalo Calves (Bubalus bubalis). J Bioanal Biomed 2: 065-068. doi:10.4172/1948-593X.1000024

Since, paracetamol reduces the doses of gentamicin and thus

expected to prevent severe toxicity in buffalo calves, the study
recommends that gentamicin can be successfully used along with
paracetamol simultaneously for treating various systemic infections
associated with pyrexia. In the present study, the mean therapeutic
concentration ( 2 g/ml) in urine was maintained upto 24 hrs and
36 hrs when gentamicin was given alone (5mg/kg i.v.) and when given
with paracetamol (40 mg/kg i.v.), respectively. Hence, the study also
recommends that gentamicin can be given by systemic route at 5mg/
kg i.v daily at every 36 hr when given along with paracetamol for
treating urinary tract infections.
Acknowledgements
Authors acknowledge Dean, B.V.C, Patna and Vice-Chancellor, R.A.U. for
providing fund for conducting this experiment.

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by

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of gentamicin in pregnant cows. Ind Vet J 66: 414-417.
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References

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rate in tissues and body fluids when it was given with paracetamol.
The t1\2 in other species were found to be low ,t1\2 of 0.05 0.01
in cow (Satish et al.,1989), 0.10.1 hrs in horse (Swan et al.,1995),
0.38 0.07 hrs rabbit (Uppal et al.,1992) and 10.251.42 min
in chicken (Garg et al.,1989) as compared to the higher t1/2 of
0.690.08 h obtained in present study. This showed that gentamicin
was distributed comparatively slowly in buffalo calves as compared to
other species noted above. Significantly higher AUC and AUMC may
be due to higher plasma drug concentrations of gentamicin obtained
from 0.042 to 0.333 hrs and 24 hrs when gentamicin was given
with paracetamol. Significantly higher value of K12 indicated faster
movement of drug from central to peripheral compartment whereas
K21 did not differ significantly in buffalo calves when gentamicin was
given alone and with paracetamol. Significantly lower Fc along with
significantly higher values of K12 and T~P showed that gentamicin
was distrbuted to a greater amount in peripheral tissues and fluids
when it was given with paracetamol. The values of and t1/2 did
not differ significantly. This indicated that similar rate of elimination
occurred in both the groups and paracetamol had no influence in
the elimination of gentamicin. Due to this MRT when gentamicin
was given alone and when given with paracetamol also did not
differ significantly. Lower Vdarea were studied by other researchers
in other species for gentamicin, Vdarea 0.370.13 L/kg in cow (Satish
et al.,1989), 0.260.4 L/Kg in goats (Garg et al.,1995), 0.45 0.11 L/
Kg in rabbit (Uppal et al.,1992) and 0.320.05 L/Kg in White Leghorn
chicken (Garg et al.,1989) as compared to present values (0.80
0.04 and 0.780.03 L/Kg) when gentamicin was given alone and
with paracetmol respectively in buffalo calves.This may indicate that
gentamicin may be distributed to be a greater amount in the body of
buffalo calves as compared to the above noted species.